Cell Signaling Technology

Product Pathways - Apoptosis / Autophagy

Phospho-Bad (Ser112) (40A9) Rabbit mAb #5284

Applications Reactivity MW (kDa) Source Isotype
W IHC-P F H Mk 23 Rabbit IgG

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  F=Flow Cytometry
Reactivity Key:  H=Human  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Bad (Ser112) (40A9) Rabbit mAb detects endogenous levels of Bad only when phosphorylated at Ser112.

Source / Purification

Phospho-Bad (Ser112) Antibody detects endogenous levels of Bad only when phosphorylated at Ser112. The Ser112 nomenclature is based upon the mouse sequence. The analogous phosphorylation site is Ser75 in human and Ser113 in rat. This antibody does not detect Bad phosphorylated at other sites, nor does it detect related family members.

Western Blotting

Western Blotting

Western blot analysis of extracts from untreated or TPA-treated COS cells, using Phospho-Bad (Ser112) (40A9) Rabbit mAb (upper), or Bad Antibody #9292 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, untreated (left) or lambda phosphatase treated (right), using Phospho-Bad (Ser 112) (40A9) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin embedded COS cells untreated (left) or TPA-treated (right), showing induced cytoplasmic staining using Phospho-Bad (Ser112) (40A9) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, using Phospho-Bad (Ser 112) (40A9) Rabbit Monoclonal Antibody preincubated with control peptide (left) or Phospho-Bad (Ser 112) Blocking Peptide (IHC Specific) #1026 (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded Non-Hodgkin's lymphoma, using Phospho-Bad (Ser112) (40A9) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of COS cells, untreated (blue) or TPA/Calyculin A treated (green), using Phospho-Bad (Ser112) (40A9) Rabbit mAb.


Background

Bad is a proapoptotic member of the Bcl-2 family that promotes cell death by displacing Bax from binding to Bcl-2 and Bcl-xL (1,2). Survival factors, such as IL-3, inhibit the apoptotic activity of Bad by activating intracellular signaling pathways that result in the phosphorylation of Bad at Ser112 and Ser136 (2). Phosphorylation at these sites promotes binding of Bad to 14-3-3 protein to prevent an association between Bad and Bcl-2 and Bcl-xl (2). Akt phosphorylates Bad at Ser136 to promote cell survival (3,4). Bad is phosphorylated at Ser112 both in vivo and in vitro by p90RSK (5,6) and mitochondria-anchored PKA (7). Phosphorylation of Ser155 in the BH3 domain by PKA plays a critical role in blocking the dimerization of Bad and Bcl-xL (8-10).

  1. Yang, E. et al. (1995) Cell 80, 285-291.
  2. Zha, J. et al. (1996) Cell 87, 619-628.
  3. Datta, S.R. et al. (1997) Cell 91, 231-241.
  4. Peso, L. et al. (1997) Science 278, 687-689.
  5. Bonni, A. et al. (1999) Science 286, 1358-1362.
  6. Tan, Y. et al. (1999) J. Biol. Chem. 274, 34859-34867.
  7. Harada, H. et al. (1999) Mol. Cell 3, 413-422.
  8. Tan, Y. et al. (2000) J. Biol. Chem. 275, 25865-25869.
  9. Lizcano, J. et al. (2000) Biochem. J. 349, 547-557.
  10. Datta, S. et al. (2000) Mol. Cell 6, 41-51.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

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